pcDNA3.1/Puro-CAG-JEDI-2P

LOCUS       pcDNA3.1/Puro-CAG-JEDI-2P                7383 bp ds-DNA     circular SYN 03-FEB-2022
DEFINITION  Genetically encoded voltage indicator (GEVI) JEDI-2P expressed under
            strong mammalian promoter (CAG).
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1/Puro-CAG-JEDI-2P
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7383)
  TITLE     Deep, rapid, and prolonged voltage imaging with an indicator 
            optimized for two-photon microscopy
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7383)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 3, 2022 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7383
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     440..459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(678..782)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        814..1193
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1472..2488
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2412..2434
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2496..2515
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2648..2657
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(4052..4069)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4058..4282
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4328..4756
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4415..4434)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4625..4646
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4765..4785)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4770..5099
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4950..5085
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5012..5031
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5110..5709
                     /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(5110..5129)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5606..5626
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5869..5990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5906..5925)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5960..5979
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6039..6055)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6039..6055)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6052..6074)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6063..6079
                     /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(6087..6117)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6132..6153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6270..6287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6441..7029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6521..6540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7200..677)
                     /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"
ORIGIN
        1 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
       61 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
      121 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
      181 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
      241 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
      301 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
      361 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
      421 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
      481 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
      541 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
      601 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
      661 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
      721 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
      781 cgcacatttc cccgaaaagt gccacctgac gtcgacattg attattgact agttattaat
      841 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      901 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      961 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     1021 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     1081 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     1141 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
     1201 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
     1261 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg gggggsgcgc
     1321 gccrggsggg gsggggsggg gsggggggsg gggsggggsg rggsggrgag gkgcggcggc
     1381 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
     1441 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt
     1501 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
     1561 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg
     1621 acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt
     1681 gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
     1741 cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct
     1801 ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg
     1861 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg
     1921 cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg agcacggccc
     1981 ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg
     2041 gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg agggctcggg
     2101 ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca
     2161 ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctgtgc
     2221 ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg cgaagcggtg
     2281 cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc
     2341 ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc gggggggacg
     2401 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
     2461 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc
     2521 tcatcatttt ggcaaagaat tcctcgagga attcactcct caggtgcagg ctgcctatca
     2581 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctgcatga
     2641 gctagccgcc accatggaga cgactgtgag gtatgaacag gggtcagagc tcactaaaac
     2701 ttcgagctct ccaacagcag atgagcccac gataaagatt gatgatggtc gtgatgaggg
     2761 taatgaacaa gacagctgtt ccaataccat taggagaaaa atttccccgt ttgtgatgtc
     2821 atttggattc agagtatttg gagttgtgct tatcattgta gacatcatag tggtgattgt
     2881 ggatctggcc atcagtgaga agaaaagagg cattagagag attcttgaag gtgtttccct
     2941 ggctatagca ctcttcttcc ttgttgatgt tctcatgaga gtgtttgttg aaggcttcaa
     3001 gaactatttc cggtccaaac tggttacttt ggatgcagtc atagtagtgg gcactctgct
     3061 aattaatatg acctactcct tctctgacct tgctgccttt aacagcgaga acgtgtatat
     3121 taccgcggat aaacagaaaa acggcattaa agcgaacttt accgtgcgcc ataacgtgga
     3181 agatggcagc gtgcagctgg cggatcatta tcagcagaac accccgattg gcgatggccc
     3241 ggtgctgctg ccggataacc attatctgag ccaccagacc gtgctgagca aagatccgaa
     3301 cgaaaaacgc gatcacatgg tgctgctgga atttgtgacc gcagcgggca ttacacacgg
     3361 catggatgaa ctgtatggcg gcaccggcgg cagcgcgagc cagggcgaag aactgtttac
     3421 cggcgtggtg ccgattctgg tggaactgga tggcgatgtg aacggccata aatttagcgt
     3481 gcgcggcgaa ggcgaaggcg atgcgaccat tggcaaactg accctgaaat ttatttgcac
     3541 caccggcaaa ctaccggtgc cgtggccgac cctggtgacc accttaacct atggcgtgca
     3601 gtgctttagc cgctatccgg atcatatgaa acgccatgat ttttttaaaa gcgcgatgcc
     3661 ggaaggctat gtgcaggaac gcaccattag ctttaaagat gatggcaaat ataaaacccg
     3721 cgcggtggtg aaatttgaag gcgataccct ggtgaaccgc attgaactga aaggcaccga
     3781 ttttaaagaa gatggcaaca ttctggggca taaactggaa tatgatacag atcagatgcc
     3841 gcacatggtt actcttcttc gagttctgaa aattgttatc ttaataagaa tatttcgcct
     3901 ggcttcacag aagaaacaac ttgaagtggt aacctaatga aagcttggta ccgagctcgg
     3961 atccactagt ccagtgtggt ggaattctgc agatatccag cacagtggcg gccgctcgag
     4021 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     4081 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4141 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4201 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4261 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4321 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4381 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4441 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4501 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4561 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4621 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4681 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4741 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4801 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4861 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4921 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4981 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5041 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5101 agctcccgca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     5161 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     5221 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     5281 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     5341 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     5401 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     5461 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5521 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     5581 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     5641 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     5701 ggtgcctgat tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5761 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5821 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5881 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5941 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     6001 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     6061 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6121 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6181 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6241 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6841 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7381 cag
//