LOCUS       Exported                6364 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Genetically encoded voltage sensor ArcLight- codon optimized.
ACCESSION   .
VERSION     .
KEYWORDS    CMV ArcLightCo (Q239)-T2A-nls-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6364)
  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 6364)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6364
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        21..1005
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     CDS             2491..2544
                     /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             2566..2586
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2587..3294
                     /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(2729..2747)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2997..3016
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3191..3214
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3387..3406)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3452..3507
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3506..3525)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3866..3882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3866..3882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3879..3901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3890..3906
                     /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(3914..3944)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3959..3980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4039..4234
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4085..4220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4147..4166
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4240..4374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4277..4296)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4331..4350
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4442..4459)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4613..5201)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4693..4712)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5372..6232)
                     /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     5995..6014
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6233..6337)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgactgca gagggccctc gaccatagcc aattcaatat ggcgtatatg gactcatgcc
       61 aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc
      121 aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc
      181 aattcaatat ggcggacttg gcactgtgcc aactggggag gggtctactt ggcacggtgc
      241 caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc
      301 caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg
      361 gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat
      421 agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca
      481 tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg
      541 tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg
      601 cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc
      661 aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct
      721 cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta
      781 ttaatagtaa cttggcaagt acattactat tggaaggacg ccagggtaca ttggcagtac
      841 tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga
      901 cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg
      961 cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaacggtac caccatggaa
     1021 ggttttgacg ggtcagactt tagccctcct gccgatctcg tgggtgtgga cggtgctgtg
     1081 atgcggaacg tggtggacgt gacaatcaac ggcgacgtga ctgcaccacc taaggcagct
     1141 cctagaaaat ctgagagtgt gaagaaagtc cactggaacg acgtggatca gggaccaagc
     1201 gagaagcccg aaacaagaca ggagaagagc aggatcacca gcgagggcga gtacatcccc
     1261 ctggaccaga tcgacatcaa cgtggacgac gggaggatgg aaattcctac cacaggagtg
     1321 ggtagagtcc agtttagggt gagagctgtc atcgaccatc tgggaatgcg ggtgttcggc
     1381 gtctttctca tcttcctgga catcattctg atgatcattg atctgagtct ccctgggaag
     1441 tcagagagct cccagagctt ctacgacggc atggcactgg ccctcagctg ctacttcatg
     1501 ctggatctcg gcctgaggat cttcgcatac ggacccaaga acttctttac caatccttgg
     1561 gaggtggccg acggactgat cattgtggtc acttttgtgg tcaccatttt ctacacagtg
     1621 ctggacgagt acgtgcagga gacaggagca gacggactcg gacagctggt ggtcctcgct
     1681 cgactgctcc gagtggtccg actggcaagg atcttttact cccaccagca ggggaattca
     1741 atgtctaaag gggaggaact gttcactggt gtggtcccca ttctggtgga gctcgacggc
     1801 gacgtgaacg gccataagtt ttcagtgagc ggggagggtg aaggcgacgc tacttacgga
     1861 aagctcaccc tgaaattcat ctgcactacc gggaagctgc cagtgccttg gcctaccctg
     1921 gtcacaactc tcacatacgg cgtgcagtgt ttcagtcgct atcctgacca tatgaagcga
     1981 cacgatttct ttaaatctgc tatgccagag ggctacgtgc aggaacggac catcttcttt
     2041 aaggacgacg gcaattataa aactcgcgca gaggtgaagt ttgaaggcga caccctggtc
     2101 aaccgaatcg agctcaaggg aattgacttc aaagaagacg gaaatatcct ggggcacaag
     2161 ctcgagtaca actataacga ccatcaggtg tacatcatgg ccgataagca gaaaaacggc
     2221 attaaggcta acttcaagat ccgccacaac attgaagacg gcggagtgca gctggccgat
     2281 cattaccagc agaacacacc aatcggagac gggcctgtcc tgctcccaga taatcactat
     2341 ctgttcacca catccactct gtctaaggac cccaacgaga aaagagatca tatggtgctg
     2401 ctggagtttg tcactgccga cgggattacc cacggaatgg acgagctgta taagttctgc
     2461 tacgagaacg aggtggcatg cggaagcgga gaaggacggg gtagtctgct gacatgcggc
     2521 gacgtagaag agaatcctgg tcccgccccg gggtcgacca tggtgcccaa gaagaagagg
     2581 aaagtcgtga gcaagggcga ggaggacaac atggccatca tcaaggagtt catgcgcttc
     2641 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     2701 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
     2761 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     2821 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2881 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2941 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     3001 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     3061 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     3121 gccgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     3181 gtcaacatca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     3241 gagcgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagctagc
     3301 agcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     3361 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     3421 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     3481 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     3541 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgccatatg ctggctgcca
     3601 tgaacaaagg tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     3661 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     3721 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     3781 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatgaagat ccctcgacct
     3841 gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3901 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3961 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4021 tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4081 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4141 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4201 aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat
     4261 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4321 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc
     4381 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4441 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4501 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4561 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4621 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4681 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4741 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4801 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4861 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4921 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4981 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5041 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5101 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5161 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5221 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5281 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5341 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5401 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5461 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5521 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5581 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5641 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5701 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5761 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5821 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5881 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5941 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6001 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6061 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6121 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6181 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6241 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6301 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6361 ctgg
//
