LOCUS       sequence-243947-        7814 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 7814)
  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 7814)
  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-Char-BFP2-TSERex
FEATURES             Location/Qualifiers
     source          1..7814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(2755..2757,2758..3468)
                     /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(2797..2818)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3667..3684)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3673..3897
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3943..4371
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4030..4049)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4240..4261
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4380..4400)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4385..4714
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4565..4700
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4627..4646
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4725..5324
                     /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(4725..4744)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5221..5241
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5484..5605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5521..5540)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5575..5594
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5654..5670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5654..5670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5667..5689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5678..5694
                     /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(5702..5708,5709..5726,5727..5732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5747..5768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5885..5902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6056..6644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6136..6155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6815..7606,7607..7675))
                     /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     7438..7457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7676..7780)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(7812..7814,1..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 gggggggggg
      541 gggggggggg gggggggcgg cggccaatca gagcggcgcg ctccgaaagt ttccttttat
      601 ggcggggcgg 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 gggggctcgg ggggggggcg 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 accatggata gcattttcga ggctgggggc
     1861 cacgggggga accctgcgcc tataaaacca attcctacaa ttgtgcctac acctacagaa
     1921 tacgagactc tgcatggtac tggccataag acactgtggg tagtattcgt gatcatgctg
     1981 atcagctcag cagtctttgc cttccgcagc tggaatatcc ctgtatcaag gcgactgtac
     2041 cacgtcatta cgactctgat tactatcacg gccgcaattt cgtattttgc tatggcctcc
     2101 ggcgacgcta cctcttttag ttgtcacagc gtggaggacc atcatggcaa acacataccg
     2161 agtactcacc acgacgtatg tagacaggtc ttctgggcac gttatgtgga ctggagcctg
     2221 accactcctc tactgctgtt agacctgagc ttactggcag gtatatcagg ggcccacacc
     2281 attatggcca ttgtcgccga cgtgatcatg gtgttggctg gattgtttgc ggcttatgga
     2341 aaggagcaca ctgcacagaa atggggctgg tacgctattg gatgtgtcgc ctacttattt
     2401 gtgatctggc acctcggtgt taatggcagg cgggcagtcg cggcgcgagg tgataaaact
     2461 accaagctgt ttagctccct tgccctgttt acactggtac tgtggacgat ttatcccatt
     2521 atttggggga tagcagatgg ggcaagaaaa gttagcgtag ataccgagat catcagctat
     2581 gccgtgctcg acgtcctggc caaacccgta ttcgggacat ggctgctgat cgctcaccgc
     2641 aatattccgg aaaccaatgt ggagttggga ggatactggg ctcacggcct aactaacggc
     2701 gaaggaagaa tccggattgg cgaagaagac gacgccgctc gaccggtagt agcagtgagc
     2761 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     2821 aacggccaca agttcagcgt gaggggcgag ggcgagggcg atgccaccaa cggcaagctg
     2881 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     2941 accctgagcc acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     3001 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3061 gacggcacct acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3121 atcgagctga agggcgtcga cttcaaggag gacggcaaca tcctggggca caagctggag
     3181 tacaacttca acagccacaa catctatatc atggccgtca agcagaagaa cggcatcaag
     3241 gtgaacttca agatccgcca caacgtggag gacggcagcg tgcagctcgc cgaccactac
     3301 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacagcca ctacctgagc
     3361 acccagtccg tgctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     3421 ttccgcaccg ccgccgggat cactctcggc atggacgagc tgtacaagaa gcttaaaagt
     3481 agaatcacaa gcgaaggcga gtacatcccc ctggatcaaa tagacataaa tgtaggtgga
     3541 ttttgttatg agaatgaatg aggtaccgag ctcggatcca ctagtccagt gtggtggaat
     3601 tctgcagata tccagcacag tggcggccgc tcgagtctag agggcccgtt taaacccgct
     3661 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3721 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3781 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3841 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3901 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3961 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4021 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4081 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4141 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4201 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4261 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4321 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     4381 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4441 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4501 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4561 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4621 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4681 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgcatgacc gagtacaagc
     4741 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     4801 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     4861 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     4921 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     4981 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     5041 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     5101 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     5161 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     5221 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     5281 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgattcgaa atgaccgacc
     5341 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5401 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5461 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5521 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5581 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5641 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5701 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5761 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5821 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5881 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5941 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6001 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6061 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6121 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6181 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6241 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6301 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6361 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6421 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6481 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6541 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6601 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6661 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6721 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6781 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6841 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6901 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6961 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7021 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7081 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7141 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7201 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7261 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7321 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7381 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7441 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7501 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7561 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7621 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7681 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7741 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7801 cacctgacgt cgac
//
