LOCUS       sequence-341196-        7910 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of voltage sensor; soma localization targeting 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7910)
  AUTHORS   Abdelfattah AS, Zheng J, Singh A, Huang YC, Reep D, Tsegaye G, Tsang
            A, Arthur BJ, Rehorova M, Olson CVL, Shuai Y, Zhang L, Fu TM, Milkie
            DE, Moya MV, Weber TD, Lemire AL, Baker CA, Falco N, Zheng Q, Grimm 
            JB, Yip MC, Walpita D, Chase M, Campagnola L, Murphy GJ, Wong AM, 
            Forest CR, Mertz J, Economo MN, Turner GC, Koyama M, Lin BJ, Betzig 
            E, Novak O, Lavis LD, Svoboda K, Korff W, Chen TW, Schreiter ER, 
            Hasseman JP, Kolb I
  TITLE     Sensitivity optimization of a rhodopsin-based fluorescent voltage 
            indicator.
  JOURNAL   Neuron. 2023 Mar 28:S0896-6273(23)00205-2. doi: 
            10.1016/j.neuron.2023.03.009.
  PUBMED    37015225
REFERENCE   2  (bases 1 to 7910)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neuron. 
            2023 Mar 28:S0896-6273(23)00205-2. doi: 
            10.1016/j.neuron.2023.03.009."
COMMENT     Sequence Label: pGP-pcDNA3.1 Puro-CAG-Voltron2-ST
FEATURES             Location/Qualifiers
     source          1..7910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      51..60
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2025..2042)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2031..2255
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2301..2729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2388..2407)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2598..2619
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2738..2758)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2743..3072
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2923..3058
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2985..3004
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3083..3682
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3083..3102)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3579..3599
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3842..3963
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3879..3898)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3933..3952
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4012..4028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4012..4028)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4025..4047)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4036..4052
                     /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)."
     promoter        complement(join(4060..4066,4067..4084,4085..4090))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4105..4126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4243..4260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4414..5002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4494..4513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5173..5964,5965..6033))
                     /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     5796..5815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6034..6138)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6170..6549
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6785..7801
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7725..7747
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7809..7828
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 tggccaatgc cctggctcac aaataccact gagatctgca tgagctagcc gccaccatgg
       61 ctgacgtgga aaccgagacc ggcatgattg cacagtggat tgtctttgct attatggctg
      121 ctgctgctat tgcttttgga gtggctgtgc actttcggcc ttcagagctg aagagcgcat
      181 actatatcaa cattgccatc tgcactatcg ccgctaccgc ttactatgca atggccgtga
      241 actaccagga cctgacaatg aatggtgaaa ggcaggtggt ctacgcaaga tatattaact
      301 gggtgctgac cacaccactg ctcctgctcg atctcatcgt catgaccaag atgggcggag
      361 tgatgatttc ttgggtcatc ggcgcagaca ttttcatgat cgtgtttggt attctgggcg
      421 acttcgagga tgaacacaag ttcaaatggg tgtactttat cgctggatgt gtgatgcagg
      481 cagtcctgac atacgggatg tataacgcca cttggaaaga cgatctgaag aaaagccccg
      541 agtaccatag ctcctatgtc agtctgctcg tcttcctgtc aatcctctgg gtgttttatc
      601 ctgtcgtgtg ggctttcggg tctggtagtg gcgtgctgtc cgtcgacaat gaggccattc
      661 tcatgggaat cctggatgtg ctcgctaagc cactgtttgg aatggggtgc ctcattgccc
      721 atgagactat cttcaagatc ggtaccggct ttccattcga cccccattat gtggaagtcc
      781 tgggcgagcg catgcactac gtcgatgttg gtccgcgcga tggcacccct gtgctgttcc
      841 tgcacggtaa cccgacctcc tcctacgtgt ggcgcaacat catcccgcat gttgcaccga
      901 cccatcgctg cattgctcca gacctgatcg gtatgggcaa atccgacaaa ccagacctgg
      961 gttatttctt cgacgaccac gtccgcttca tggatgcctt catcgaagcc ctgggtctgg
     1021 aagaggtcgt cctggtcatt cacgactggg gctccgctct gggtttccac tgggccaagc
     1081 gcaatccaga gcgcgtcaaa ggtattgcat ttatggagtt catccaccct atcccgacct
     1141 gggacgaatg gccagaattt gcccgcgaga ccttccaggc cttccgcacc accgacgtcg
     1201 gccgcaagct gatcatcgat cagaacgttt ttatcgaggg tacgctgccg atgggtgtcg
     1261 tccgcccgct gactgaagtc gagatggacc attaccgcga gccgttcctg aatcctgttg
     1321 accgcgagcc actgtggcgc ttcccaaacg agctgccaat cgccggtgag ccagcgaaca
     1381 tcgtcgcgct ggtcgaagaa tacatggact ggctgcacca gtcccctgtc ccgaagctgc
     1441 tgttctgggg caccccaggc gttctgatcc caccggccga agccgctcgc ctggccaaaa
     1501 gcctgcctaa ctgcaaggct gtggacatcg gcccgggtct gaatctgctg caagaagaca
     1561 acccggacct gatcggcagc gagatcgcgc gctggctgtc gacgctcgag atttccggcg
     1621 agccaaccac taagagcagg atcaccagcg agggcgagta catccccctg gaccagatcg
     1681 acatcaacgt gttctgctac gagaacgagg tgcaaagtca gcctatcctg aacacaaagg
     1741 aaatggctcc acagtctaag cctcccgaag agcttgagat gtccagtatg ccaagtcccg
     1801 tggctcccct ccctgccagg actgaaggag tgattgacat gaggagtatg tcatctattg
     1861 atagcttcat ctcttgcgca acagatttcc ccgaggctac tcgattctaa tgaaagcttg
     1921 ctaccgagct cggatccact agtccagtgt ggtggaattc tgcagatatc cagcacagtg
     1981 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     2041 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2101 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2161 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2221 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2281 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2341 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2401 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2461 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2521 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2581 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2641 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2701 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     2761 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2821 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2881 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2941 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3001 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3061 ggcttttgca aaaagctccc gcatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3121 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3181 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3241 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     3301 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     3361 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     3421 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     3481 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     3541 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     3601 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     3661 gacccgcaag cccggtgcct gattcgaaat gaccgaccaa gcgacgccca acctgccatc
     3721 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3781 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     3841 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3901 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3961 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4021 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4081 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4141 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4201 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4261 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4321 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4381 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4441 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4501 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4561 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4621 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4681 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4741 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4801 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4861 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4921 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4981 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5041 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5101 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5161 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5221 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5281 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5341 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5401 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5461 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5521 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5581 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5641 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5701 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5761 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5821 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5881 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5941 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6001 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6061 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6121 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acattgatta
     6181 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     6241 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     6301 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     6361 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     6421 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     6481 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     6541 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     6601 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
     6661 gggggggggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
     6721 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     6781 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
     6841 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
     6901 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
     6961 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
     7021 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
     7081 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
     7141 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
     7201 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
     7261 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
     7321 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     7381 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc
     7441 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
     7501 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
     7561 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
     7621 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
     7681 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     7741 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     7801 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcctcga
     7861 ggaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
//
