LOCUS       Exported                5507 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression vector for expression of DsRed.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-DsRed
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5507)
  AUTHORS   Matsuda T, Cepko CL
  TITLE     Electroporation and RNA interference in the rodent retina in vivo 
            and in vitro.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22.
  PUBMED    14603031
REFERENCE   2  (bases 1 to 5507)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5507
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1763..1772
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1769..2446
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(1949..1969)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2361..2384
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2531..2550)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2596..2651
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2650..2669)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3010..3026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3010..3026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3023..3045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3034..3050
                     /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(3058..3088)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3103..3124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3183..3378
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3229..3364
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3291..3310
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3384..3518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3421..3440)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3475..3494
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3586..3603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3757..4345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3837..3856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4516..5376)
                     /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     5139..5158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5377..5481)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attctgcagt cgacggtacc
     1741 gcgggcccgg gatccaccgg tcgccaccat ggcctcctcc gagaacgtca tcaccgagtt
     1801 catgcgcttc aaggtgcgca tggagggcac cgtgaacggc cacgagttcg agatcgaggg
     1861 cgagggcgag ggccgcccct acgagggcca caacaccgtg aagctgaagg tgaccaaggg
     1921 cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg gctccaaggt
     1981 gtacgtgaag caccccgccg acatccccga ctacaagaag ctgtccttcc ccgagggctt
     2041 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gcgaccgtga cccaggactc
     2101 ctccctgcag gacggctgct tcatctacaa ggtgaagttc atcggcgtga acttcccctc
     2161 cgacggcccc gtgatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta
     2221 cccccgcgac ggcgtgctga agggcgagac ccacaaggcc ctgaagctga aggacggcgg
     2281 ccactacctg gtggagttca agtccatcta catggccaag aagcccgtgc agctgcccgg
     2341 ctactactac gtggacgcca agctggacat cacctcccac aacgaggact acaccatcgt
     2401 ggagcagtac gagcgcaccg agggccgcca ccacctgttc ctgtagcggc cgcactcctc
     2461 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     2521 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     2581 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     2641 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     2701 atttggttta gagtttggca acatatgcca tatgctggct gccatgaaca aaggtggcta
     2761 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     2821 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     2881 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     2941 ccagtcatag ctgtccctct tctcttatga agatccctcg acctgcagcc caagcttggc
     3001 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3061 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3121 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     3181 ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3241 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3301 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3361 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa
     3421 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3481 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca
     3541 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3601 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3661 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3721 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3781 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3841 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3901 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     3961 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4021 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4081 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4141 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4201 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4261 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4321 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4381 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4441 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4501 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4561 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4621 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4681 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4741 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4801 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4861 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4921 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4981 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5041 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5101 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5161 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5221 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5281 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5341 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5401 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5461 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
