LOCUS       Exported                7815 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses the proton pump rhodopsin in mammalian cells under CAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1-CarO-BFP2-TSERex
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7815)
  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 7815)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7815
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     regulatory      1829..1838
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2756..3469
                     /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(2798..2819)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3668..3685)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3674..3898
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3944..4372
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4031..4050)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4241..4262
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4381..4401)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4386..4715
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4566..4701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4628..4647
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4726..5325
                     /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(4726..4745)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5222..5242
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5485..5606
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5522..5541)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5576..5595
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5655..5671)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5655..5671)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5668..5690)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5679..5695
                     /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(5703..5733)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5748..5769
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5886..5903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6057..6645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6137..6156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6816..7676)
                     /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     7439..7458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7677..7781)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7813..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 cgccgggcgg gggggggggg gggggggggg gggggggggg
      541 ggggcgggga ggggcggcgg 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 caccatggca gaccacttat atgcccgcaa
     1861 gaatgacgcc ctcaatgtta atccagatat cgttaatggt caacgttctg acattaatat
     1921 cacagtgagg ggctctgatt ggtattgggc tgtgtgtgca gtgatgactg tgagcacctt
     1981 cgcatttctg gggttgggaa tgcggaaacc gagaacagat aggatctttc actacatcac
     2041 ggcaggcatt acaatgatcg cctctatagc ttacttcacc atggcttcca acctcggttg
     2101 gactcctatc gcagtggaat ttcaacgatc aaaccaccgt gtggctggga tttacaggga
     2161 gatcttctac gcccgctata tagactggtt tctcactacc cccctgctcc ttaccgattt
     2221 gctgctgacc gcgggaatgc cctggccgac tgttttgtgg gttatcttgg tggactgggt
     2281 aatgatcgtg acggggttgg tgggcgcact cgttaagtct agttacaagt ggggctattt
     2341 tgcttttggc tgcgccgccc tggcatacat cgtgtatgtg ctagcatggg aggcccgtct
     2401 gcacgccaaa cacgttggac ctgatgtggg caggactttt gtcatgtgtg ggtccctgac
     2461 cgcagtagtg tggatactgt accctatcgc ttggggcgtg tgcgagggtg gaaacctcat
     2521 agctcctgat tcagaggcag tgttttatgg catcttggat ctgatagcaa agcccgtgtt
     2581 tggagccctg ttactgtggg gtcacagaaa catcgaccca gcccgcctgg ggcttaggat
     2641 tagagatatt gatgagcgca tttttccaga cgggccaaat aataaggttg caagcgggca
     2701 cggggcccgg aacgacaccg ccaccgcaag cggatcaaat gtcaacccta atgccgtgag
     2761 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2821 aaacggccac aagttcagcg tgaggggcga gggcgagggc gatgccacca acggcaagct
     2881 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2941 caccctgagc cacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     3001 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3061 cgacggcacc tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3121 catcgagctg aagggcgtcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3181 gtacaacttc aacagccaca acatctatat catggccgtc aagcagaaga acggcatcaa
     3241 ggtgaacttc aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta
     3301 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacagcc actacctgag
     3361 cacccagtcc gtgctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3421 gttccgcacc gccgccggga tcactctcgg catggacgag ctgtacaaga aaagtagaat
     3481 cacaagcgaa ggcgagtaca tccccctgga tcaaatagac ataaatgtag gtggattttg
     3541 ttatgagaat gaatgaaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
     3601 ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc
     3661 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3721 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3781 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3841 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3901 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3961 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4021 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4081 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4141 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4201 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4261 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4321 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4381 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4441 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4501 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4561 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4621 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4681 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgcatgac cgagtacaag
     4741 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     4801 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     4861 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4921 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4981 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     5041 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     5101 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     5161 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     5221 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     5281 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgattcga aatgaccgac
     5341 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5401 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5461 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5521 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5581 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5641 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5701 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5761 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5821 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5881 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5941 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6001 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6061 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6481 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6901 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6961 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7021 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7081 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7141 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7201 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7261 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7321 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7381 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7441 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7501 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7561 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7621 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7681 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7741 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7801 ccacctgacg tcgac
//
