LOCUS       sequence-240926-        7773 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 7773)
  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 7773)
  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-Lari-BFP2-TSERex
FEATURES             Location/Qualifiers
     source          1..7773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(890..892,893..1603)
                     /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(932..953)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1802..1819)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1808..2032
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2078..2506
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2165..2184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2375..2396
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2515..2535)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2520..2849
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2700..2835
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2762..2781
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2860..3459
                     /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(2860..2879)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3356..3376
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3619..3740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3656..3675)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3710..3729
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3789..3805)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3789..3805)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3802..3824)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3813..3829
                     /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(3837..3843,3844..3861,3862..3867))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3882..3903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4020..4037)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4191..4779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4271..4290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4950..5741,5742..5810))
                     /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     5573..5592
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5811..5915)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5947..6326
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6605..7621
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7545..7567
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7629..7648
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 gctagccgcc accatgctta acacgcagtt gcagaacctg cttaataatc ctctgacccc
       61 catccaggct tatgctgatt cagacctgtt ctcaactcag aatgtcctct cccggctcag
      121 ccactcagtg ctctgggttg tgtttgccat aatgattctc agcactgtcg tcataggaat
      181 cctgacaacc cagcagcaga agaataataa actgtttcac cacatcactc taaccatcaa
      241 ctttattgcc gcgatcagct actactctat ggctagtggc attggtatct ctttatcaca
      301 ccacggcgcg cgacctttat tctgggcccg gtacttggac tggttactga ctaccccact
      361 tctcctaacc gatttgggcc tcctggccgg aatacccatt caggacatcg tgctgcttgt
      421 gctgtcagat gtagtgatgg tagtcactgg cttgctcggc gctatcaacc agagcgaatc
      481 aactaagtgg ggctactttg tgatgtcatg cattttcttt gtttacgtgg tctatgaact
      541 ggcgaccacc gcaagaagaa cgtcctatct gaaaagttca agagtcggct tcttgtttac
      601 agctatcgca gtgtactcca tgctgctttg gactgtgtac cctgtggtgt gggctcttag
      661 cgaaggctca agtatcatca gtcctgatct ggaaatactg ttttatgctg tgttggacgt
      721 gatggcaaag gctgtgttcg gcttttggct tctaatcgcc cacagcatca ttccagaatc
      781 aaatgtgatt gtttccaata gctttaccga cccccgaata cagaacggag tcggatattc
      841 cgccatcgcc gagaatgaag aagccgtgag ggaccgaccg gtagtagcag tgagcaaggg
      901 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      961 ccacaagttc agcgtgaggg gcgagggcga gggcgatgcc accaacggca agctgaccct
     1021 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1081 gagccacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     1141 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1201 cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1261 gctgaagggc gtcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1321 cttcaacagc cacaacatct atatcatggc cgtcaagcag aagaacggca tcaaagtgaa
     1381 cttcaagatc cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc actaccagca
     1441 gaacaccccc atcggcgacg gccccgtgct gctgcccgac agccactacc tgagcaccca
     1501 gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttccg
     1561 caccgccgcc gggattactc tcggcatgga cgagctgtac aagaagctta aaagtagaat
     1621 cacaagcgaa ggcgagtaca tccccctgga tcaaatagac ataaatgtag gtggattttg
     1681 ttatgagaat gaatgaggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
     1741 agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca
     1801 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1861 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     1921 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     1981 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2041 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2101 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2161 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2221 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2281 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2341 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2401 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2461 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2521 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2581 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2641 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2701 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2761 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     2821 gcttttttgg aggcctaggc ttttgcaaaa agctcccgca tgaccgagta caagcccacg
     2881 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     2941 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     3001 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     3061 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     3121 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     3181 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     3241 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     3301 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     3361 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     3421 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgat tcgaaatgac cgaccaagcg
     3481 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     3541 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     3601 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3661 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3721 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     3781 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3841 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3901 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3961 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4021 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4081 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4141 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4201 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4261 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4321 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4381 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4441 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4501 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4561 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4621 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4681 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4741 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4801 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4861 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4921 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4981 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5041 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5101 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5161 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5221 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5281 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5341 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5401 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5461 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5521 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5581 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5641 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5701 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5761 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5821 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5881 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5941 gacgtcgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     6001 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     6061 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     6121 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     6181 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     6241 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     6301 acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc
     6361 ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg
     6421 cagcgatggg ggcggggggg gggggnnnsg cgcgccrggc ggggsggggs ggggsgrggg
     6481 gsggggsggg gsgrggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
     6541 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     6601 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
     6661 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
     6721 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
     6781 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
     6841 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
     6901 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
     6961 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
     7021 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
     7081 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
     7141 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     7201 ggcggggccg cctcgggccg gggrgggctc gggggagggg cgcggcggcc cccggagcgc
     7261 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
     7321 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
     7381 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
     7441 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
     7501 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     7561 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     7621 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcctcga
     7681 ggaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     7741 cctggctcac aaataccact gagatctgca tga
//
