LOCUS       sequence-334519-        8367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresses SCN2B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8367)
  TITLE     These plasmids allow lentivirus-mediated overexpression of 
            understudied V5 tagged ion channels in mammalian cell lines.
REFERENCE   2  (bases 1 to 8367)
  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_SCN2B_OE_1
FEATURES             Location/Qualifiers
     source          1..8367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..46
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    163..184
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(199..204,205..222,223..229)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    237..253
                     /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     242..264
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     261..277
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     261..277
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     296..316
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        298..316
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        344..570
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             571..751
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    798..923
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1416..1649
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1834..1878
                     /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        2053..2553
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2472..2490
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2565..2960
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    3020..3137
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3254..3557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3558..3761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3712..3732
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    3784..3808
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4454..4478)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4480..4521
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    4563..5151
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4616..4636)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5034..5045)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5223..5456
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5534..5655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5571..5590)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5625..5644
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5695..5830
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5850..5869)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5851..5869)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5879..5896)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5879..5895)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5888..5910)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6037..6492
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6124..6143)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6334..6355
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6518..6622
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6623..6691,6692..7483)
                     /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(6841..6860)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7654..8242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8143..8162
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
       61 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      121 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
      181 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
      241 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
      301 taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata
      361 ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa
      421 aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg
      481 caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt
      541 gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc
      601 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
      661 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
      721 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
      781 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
      841 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
      901 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
      961 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     1021 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     1081 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     1141 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     1201 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     1261 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     1321 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     1381 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     1441 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     1501 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     1561 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     1621 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     1681 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     1741 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     1801 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     1861 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     1921 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     1981 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     2041 ccgaggggac ccttgcgcct tttccaaggc agccctgggt ttgcgcaggg acgcggctgc
     2101 tctgggcgtg gttccgggaa acgcagcggc gccgaccctg ggtctcgcac attcttcacg
     2161 tccgttcgca gcgtcacccg gatcttcgcc gctacccttg tgggcccccc ggcgacgctt
     2221 cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa
     2281 acggaagccg cacgtctcac tagtaccctc gcagacggac agcgccaggg agcaatggca
     2341 gcgcgccgac cgcgatgggc tgtggccaat agcggctgct cagcagggcg cgccgagagc
     2401 agcggccggg aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc
     2461 ctgcccgcgc ggtgttccgc attctgcaag cctccggagc gcacgtcggc agtcggctcc
     2521 ctcgttgacc gaatcaccga cctctctccc cagggggtac caccatggcc aagcctttgt
     2581 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     2641 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     2701 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     2761 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     2821 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     2881 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     2941 ctggttatgt gtgggagggc ctgcagctgc agtagtaaga attctagatc ttgagacaaa
     3001 tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg
     3061 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
     3121 caaaaattca aaattttcgg gtttattaca gggacagcag agatccactt tggcgccggc
     3181 tcgagggggc ccggggttat taatagtaat caattacggg gtcattagtt catagcccat
     3241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     3301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     3361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     3421 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     3481 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     3541 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     3601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     3661 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     3721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actgtcggga
     3781 tcaacaagtt tgtacaaaaa agttggcatg cacagagatg cctggctacc tcgccctgcc
     3841 ttcagcctca cggggctcag tctctttttc tctttggtgc caccaggacg gagcatggag
     3901 gtcacagtac ctgccaccct caacgtcctc aatggctctg acgcccgcct gccctgcacc
     3961 ttcaactcct gctacacagt gaaccacaaa cagttctccc tgaactggac ttaccaggag
     4021 tgcaacaact gctctgagga gatgttcctc cagttccgca tgaagatcat taacctgaag
     4081 ctggagcggt ttcaagaccg cgtggagttc tcagggaacc ccagcaagta cgatgtgtcg
     4141 gtgatgctga gaaacgtgca gccggaggat gaggggattt acaactgcta catcatgaac
     4201 ccccctgacc gccaccgtgg ccatggcaag atccatctgc aggtcctcat ggaagagccc
     4261 cctgagcggg actccacggt ggccgtgatt gtgggtgcct ccgtcggggg cttcctggct
     4321 gtggtcatct tggtgctgat ggtggtcaag tgtgtgagga gaaaaaaaga gcagaagctg
     4381 agcacagatg acctgaagac cgaggaggag ggcaagacgg acggtgaagg caacccggat
     4441 gatggcgcca agtacccaac tttcttgtac aaagtggttg gtaagcctat ccctaaccct
     4501 ctcctcggtc tcgattctac gtagtaatga gctagcgcta accggtggcg cgttaagtcg
     4561 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     4621 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4681 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     4741 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     4801 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     4861 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     4921 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     4981 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5041 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5101 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt
     5161 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     5221 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
     5281 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     5341 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     5401 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtac
     5461 gtatagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     5521 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5581 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5641 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg
     5701 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5761 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5821 tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     5881 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     5941 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     6001 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     6061 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     6121 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     6181 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     6241 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     6301 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     6361 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     6421 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     6481 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6541 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6601 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6661 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6721 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6781 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6841 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6901 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6961 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7021 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7081 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7141 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7201 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7261 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7321 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7381 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7441 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7501 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7561 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7621 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7681 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7741 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7801 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7861 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7921 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7981 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8041 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8101 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8161 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8221 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     8281 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     8341 tattaccgcc tttgagtgag ctgatac
//
