LOCUS       sequence-243946-        7848 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 7848)
  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 7848)
  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-Mac-BFP2-TSERex
FEATURES             Location/Qualifiers
     source          1..7848
                     /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(2789..2791,2792..3502)
                     /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(2831..2852)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3701..3718)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3707..3931
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3977..4405
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4064..4083)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4274..4295
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4414..4434)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4419..4748
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4599..4734
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4661..4680
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4759..5358
                     /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(4759..4778)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5255..5275
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5518..5639
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5555..5574)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5609..5628
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5688..5704)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5688..5704)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5701..5723)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5712..5728
                     /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(5736..5742,5743..5760,5761..5766))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5781..5802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5919..5936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6090..6678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6170..6189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6849..7640,7641..7709))
                     /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     7472..7491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7710..7814)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(7846..7848,1..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 ggcggggggg 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 caccatgatc gtggaccagt tcgaggaggt
     1861 gctgatgaag accagccagc tgttcccact gccaaccgct acccagagcg cccagccaac
     1921 ccacgtggcc cccgtgccaa ccgtgctgcc cgacaccccc atctacgaga ccgtgggcga
     1981 cagcggcagc aagaccctgt gggtggtgtt cgtgctgatg ctgatcgcca gcgccgcctt
     2041 caccgccctg agctggaaga tccccgtgaa caggaggctg taccacgtga tcaccaccat
     2101 catcaccctg accgccgccc tgagctactt cgctatggct accggccacg gagtggccct
     2161 gaacaagatc gtgatcagga cccagcacga ccacgtgccc gacacctacg agaccgtgta
     2221 ccgacaggtg tactacgcca ggtacatcga ctgggctatc accaccccac tgctgctgct
     2281 ggacctggga ctgctggctg gaatgagcgg agcccacatc ttcatggcca tcgtggctga
     2341 cctgatcatg gtgctgaccg gcctgttcgc tgctttcggc agcgagggaa ccccacagaa
     2401 gtggggatgg tacaccatcg cctgcatcgc ctacatcttc gtggtgtggc acctggtgct
     2461 gaacggcggc gccaacgcca gggtgaaggg cgagaagctg aggagcttct tcgtggccat
     2521 cggagcttac accctgatcc tgtggaccgc ttacccaatc gtgtggggac tggctgacgg
     2581 agctaggaag atcggagtgg acggagagat catcgcttac gctgtgctgg acgtgctggc
     2641 taagggagtg ttcggagctt ggctgctggt gacccacgcc aacctgaggg agagcgacgt
     2701 ggagctgaac ggcttctggg ccaacggcct gaacagggag ggcgccatca ggatcggcga
     2761 ggacgacggc gcccgaccgg tagtagcagt gagcaagggc gaggagctgt tcaccggggt
     2821 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgagggg
     2881 cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg
     2941 caagctgccc gtgccctggc ccaccctcgt gaccaccctg agccacggcg tgcagtgctt
     3001 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     3061 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc acctacaaga cccgcgccga
     3121 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggcg tcgacttcaa
     3181 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacatcta
     3241 tatcatggcc gtcaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacgt
     3301 ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3361 ccccgtgctg ctgcccgaca gccactacct gagcacccag tccgtgctga gcaaagaccc
     3421 caacgagaag cgcgatcaca tggtcctgct ggagttccgc accgccgccg ggatcactct
     3481 cggcatggac gagctgtaca agaagcttaa aagtagaatc acaagcgaag gcgagtacat
     3541 ccccctggat caaatagaca taaatgtagg tggattttgt tatgagaatg aatgaggtac
     3601 cgagctcgga tccactagtc cagtgtggtg gaattctgca gatatccagc acagtggcgg
     3661 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3721 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3781 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3841 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3901 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3961 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4021 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4081 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4141 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4201 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4261 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4321 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4381 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4441 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4501 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4561 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4621 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4681 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4741 tttgcaaaaa gctcccgcat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     4801 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     4861 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     4921 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     4981 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     5041 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     5101 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     5161 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     5221 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     5281 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     5341 cgcaagcccg gtgcctgatt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5401 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5461 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5521 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5581 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5641 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5701 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5761 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5821 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5881 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5941 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6001 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6061 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6121 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6181 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6241 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6301 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6361 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6421 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6481 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6541 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6601 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6661 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6721 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6781 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6841 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6901 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6961 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7021 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7081 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7141 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7201 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7261 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7321 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7381 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7441 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7501 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7561 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7621 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7681 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7741 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7801 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgac
//
