LOCUS       sequence-243944-        7818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the proton pump rhodopsin in mammalian cells under CAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7818)
  AUTHORS   Wu L, Dong A, Dong L, Wang SQ, Li Y
  TITLE     PARIS, an optogenetic method for functionally mapping gap junctions.
  JOURNAL   Elife. 2019 Jan 14;8. pii: 43366. doi: 10.7554/eLife.43366.
  PUBMED    30638447
REFERENCE   2  (bases 1 to 7818)
  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"; journalName: "Elife."; 
            date: "2019-01-14"; volume: "8. pii: 43366. doi"; pages: " 
            10.7554/eLife.43366"
COMMENT     Sequence Label: pcDNA3.1-Nod-BFP2-TSERex
FEATURES             Location/Qualifiers
     source          1..7818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        379..652
                     /label=chicken beta-actin promoter
     intron          653..1669
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1593..1615
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1677..1696
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(2759..2761,2762..3472)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIKV
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLLE
                     FRTAAGITLGMDELYK"
     primer_bind     complement(2801..2822)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3671..3688)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3677..3901
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3947..4375
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4034..4053)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4244..4265
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4384..4404)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4389..4718
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4569..4704
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4631..4650
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4729..5328
                     /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(4729..4748)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5225..5245
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5488..5609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5525..5544)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5579..5598
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5658..5674)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5658..5674)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5671..5693)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5682..5698
                     /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(5706..5712,5713..5730,5731..5736))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5751..5772
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5889..5906)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6060..6648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6140..6159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6819..7610,7611..7679))
                     /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     7442..7461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7680..7784)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(7816..7818,1..377)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
       61 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      121 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      181 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      241 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      301 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      361 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
      421 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
      481 gggcgggggg ggggggcgcg cgccaggcgg gggggggggg gggggggggg gggggggggc
      541 ggggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
      601 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
      661 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
      721 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      781 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga
      841 ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt
      901 gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg
      961 cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc
     1021 cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg
     1081 ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc
     1141 ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg
     1201 gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc
     1261 tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg
     1321 aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact
     1381 tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag
     1441 cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg
     1501 tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga
     1561 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     1621 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     1681 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctcgagg aattcactcc
     1741 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     1801 ataccactga gatctgcatg agctagccgc caccatgttg gtggaacagt tcgaagaggt
     1861 cctgaagaaa actagcggac tgcctttgcc tactgcctca cctacccatg tcgctccaat
     1921 tccatctgtc gtacccacaa ccccagagct ccagtttatt ggggagagcg gtcagaaaac
     1981 actgtgggtg gtgttcgtcc tcatgattat tgcatccgca ggcttcaccg ctttaagctg
     2041 gcgggtgcct ttatccaaga gactgtatca caccatcacc accatgatca ccatttttgc
     2101 tgccctgtca tatttcgcca tggctaccgg tcacggcgta agtgtgcaga agatcattgt
     2161 acgggagcaa cacgaccacg tccccgatac atttacagaa gttcataggc aggttttctg
     2221 ggctaggtac gtcgattggt ccgtaacgac cccactcctc ctgttggact tgggattgct
     2281 ggcagggatg tctggcggcc atattatcat ggccatcgtg gccgacctta taatgatctt
     2341 aactggtctg ttcgccgcat tcggcgagga aggtacacct cagaagtggg gatggtacac
     2401 catcgcctgc atcgcctata tttttgtcat ttggcatctt gctctgaacg gcggcgcaaa
     2461 cgcaacgagc aagggcccga agcttaggag ctttttcgtc gcaatcggag gttacactct
     2521 cctcctttgg actgcctacc caatagtgtg gggcctggct gacggtagtc ggaagattgg
     2581 cgtcgatggg gaagtcatcg cctatgcaat ccttgacgta ttggcaaagg gcgtgtttgg
     2641 agcctggtta ctgattaccc acgctaaact gcgcgagagc gacgtagagt tgaacggatt
     2701 ttggagcaac ggattaaata gcgaaggggc cgtgcgcctg ggcgaggacg acggtgcggt
     2761 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2821 cgtaaacggc cacaagttca gcgtgagggg cgagggcgag ggcgatgcca ccaacggcaa
     2881 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2941 gaccaccctg agccacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     3001 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3061 ggacgacggc acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3121 ccgcatcgag ctgaagggcg tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3181 ggagtacaac ttcaacagcc acaacatcta tatcatggcc gtcaagcaga agaacggcat
     3241 caaggtgaac ttcaagatcc gccacaacgt ggaggacggc agcgtgcagc tcgccgacca
     3301 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca gccactacct
     3361 gagcacccag tccgtgctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3421 ggagttccgc accgccgccg ggatcactct cggcatggac gagctgtaca agaaaagtag
     3481 aatcacaagc gaaggcgagt acatccccct ggatcaaata gacataaatg taggtggatt
     3541 ttgttatgag aatgaatgaa agcttggtac cgagctcgga tccactagtc cagtgtggtg
     3601 gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     3661 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3721 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3781 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3841 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3901 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3961 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4021 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4081 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4141 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4201 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4261 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4321 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4381 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4441 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4501 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4561 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4621 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4681 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccgcat gaccgagtac
     4741 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     4801 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     4861 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     4921 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     4981 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     5041 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     5101 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     5161 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     5221 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     5281 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatt cgaaatgacc
     5341 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5401 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5461 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5521 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5581 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5641 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5701 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5761 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5821 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5881 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5941 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6001 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6061 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6121 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6181 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6241 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6301 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6361 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6421 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6481 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6541 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6601 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6661 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6721 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6781 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6841 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6901 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6961 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7021 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7081 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7141 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7201 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7261 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7321 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7381 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7441 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7501 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7561 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7621 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7681 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7741 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7801 gtgccacctg acgtcgac
//
