LOCUS       Exported                5835 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Genetically encoded voltage sensor ArcLight- codon optimized.
ACCESSION   .
VERSION     .
KEYWORDS    hSyn ArcLightCo (Q239)-T2A-nls-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5835)
  AUTHORS   Platisa J, Vasan G, Yang A, Pieribone VA
  TITLE     Directed evolution of key residues in fluorescent protein inverses 
            the polarity of voltage sensitivity in the genetically-encoded 
            indicator ArcLight.
  JOURNAL   ACS Chem Neurosci. 2017 Jan 3. doi: 10.1021/acschemneuro.6b00234.
  PUBMED    28045247
REFERENCE   2  (bases 1 to 5835)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5835
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(68..84)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(68..84)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(81..103)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    92..108
                     /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(116..146)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    161..182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        241..436
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      287..422
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     349..368
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    442..576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(479..498)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     533..552
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(644..661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(815..1403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(895..914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1574..2434)
                     /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     2197..2216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2435..2539)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2593..3040
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             4529..4582
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4604..4624
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4625..5332
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(4767..4785)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5035..5054
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5229..5252
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(5422..5441)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5487..5542
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5541..5560)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
ORIGIN
        1 agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt
       61 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      121 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
      181 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc
      241 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
      301 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
      361 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
      421 cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc
      481 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
      541 tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac
      601 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
      661 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      721 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      781 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      841 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      901 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      961 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1021 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1081 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1141 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1201 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1261 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1321 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1381 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1441 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1501 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1561 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1621 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1681 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1741 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1801 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1861 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1921 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1981 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2041 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2101 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2161 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2221 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2281 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2341 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2401 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2461 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2521 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcg actgcagagg
     2581 gccctgcgta tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta
     2641 cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca
     2701 tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc
     2761 ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag
     2821 cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct
     2881 tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg
     2941 ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc
     3001 ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag accggtacca ccatggaagg
     3061 ttttgacggg tcagacttta gccctcctgc cgatctcgtg ggtgtggacg gtgctgtgat
     3121 gcggaacgtg gtggacgtga caatcaacgg cgacgtgact gcaccaccta aggcagctcc
     3181 tagaaaatct gagagtgtga agaaagtcca ctggaacgac gtggatcagg gaccaagcga
     3241 gaagcccgaa acaagacagg agaagagcag gatcaccagc gagggcgagt acatccccct
     3301 ggaccagatc gacatcaacg tggacgacgg gaggatggaa attcctacca caggagtggg
     3361 tagagtccag tttagggtga gagctgtcat cgaccatctg ggaatgcggg tgttcggcgt
     3421 ctttctcatc ttcctggaca tcattctgat gatcattgat ctgagtctcc ctgggaagtc
     3481 agagagctcc cagagcttct acgacggcat ggcactggcc ctcagctgct acttcatgct
     3541 ggatctcggc ctgaggatct tcgcatacgg acccaagaac ttctttacca atccttggga
     3601 ggtggccgac ggactgatca ttgtggtcac ttttgtggtc accattttct acacagtgct
     3661 ggacgagtac gtgcaggaga caggagcaga cggactcgga cagctggtgg tcctcgctcg
     3721 actgctccga gtggtccgac tggcaaggat cttttactcc caccagcagg ggaattcaat
     3781 gtctaaaggg gaggaactgt tcactggtgt ggtccccatt ctggtggagc tcgacggcga
     3841 cgtgaacggc cataagtttt cagtgagcgg ggagggtgaa ggcgacgcta cttacggaaa
     3901 gctcaccctg aaattcatct gcactaccgg gaagctgcca gtgccttggc ctaccctggt
     3961 cacaactctc acatacggcg tgcagtgttt cagtcgctat cctgaccata tgaagcgaca
     4021 cgatttcttt aaatctgcta tgccagaggg ctacgtgcag gaacggacca tcttctttaa
     4081 ggacgacggc aattataaaa ctcgcgcaga ggtgaagttt gaaggcgaca ccctggtcaa
     4141 ccgaatcgag ctcaagggaa ttgacttcaa agaagacgga aatatcctgg ggcacaagct
     4201 cgagtacaac tataacgacc atcaggtgta catcatggcc gataagcaga aaaacggcat
     4261 taaggctaac ttcaagatcc gccacaacat tgaagacggc ggagtgcagc tggccgatca
     4321 ttaccagcag aacacaccaa tcggagacgg gcctgtcctg ctcccagata atcactatct
     4381 gttcaccaca tccactctgt ctaaggaccc caacgagaaa agagatcata tggtgctgct
     4441 ggagtttgtc actgccgacg ggattaccca cggaatggac gagctgtata agttctgcta
     4501 cgagaacgag gtggcatgcg gaagcggaga aggacggggt agtctgctga catgcggcga
     4561 cgtagaagag aatcctggtc ccgccccggg gtcgaccatg gtgcccaaga agaagaggaa
     4621 agtcgtgagc aagggcgagg aggacaacat ggccatcatc aaggagttca tgcgcttcaa
     4681 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     4741 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc
     4801 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     4861 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     4921 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     4981 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     5041 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     5101 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     5161 cgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     5221 caacatcaag ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     5281 gcgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aagctagcgg
     5341 ccgcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct
     5401 ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa
     5461 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     5521 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     5581 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
     5641 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     5701 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     5761 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     5821 ctcctgacta ctccc
//
