LOCUS       Exported                5541 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  mammalian expression of tdTomato controlled by the ubiquitously 
            active CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    pCSCMV:tdTomato
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5541)
  AUTHORS   Waldner C, Roose M, Ryffel GU
  TITLE     Red fluorescent Xenopus laevis: a new tool for grafting analysis.
  JOURNAL   BMC Dev Biol. 2009  . 9():37.
  PUBMED    19549299
REFERENCE   2  (bases 1 to 5541)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5541
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1064..1073
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1070..2500
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     primer_bind     complement(2521..2537)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2522..2538)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2549..2566)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2571..2705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2595..2614)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2649..2668
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2818..2838)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2818..2836)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2857..2873)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2857..2873)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2870..2892)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2881..2897
                     /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(2905..2935)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2950..2971
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3088..3105)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3259..3847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3339..3358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4018..4878)
                     /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     4641..4660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4879..4983)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5009..5464)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5146..5167)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5358..5377
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat cccgccacca tggtgagcaa
     1081 gggcgaggag gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa
     1141 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     1201 cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc
     1261 ccagttcatg tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa
     1321 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     1381 tctggtgacc gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa
     1441 gatgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg
     1501 ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca
     1561 ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc
     1621 caagaagccc gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc
     1681 ccacaacgag gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct
     1741 gttcctgggg catggcaccg gcagcaccgg cagcggcagc tccggcaccg cctcctccga
     1801 ggacaacaac atggccgtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc
     1861 catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1921 ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct
     1981 gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga
     2041 ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     2101 cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa
     2161 ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat
     2221 gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat
     2281 ccaccaggcc ctgaagctga aggacggcgg ccgctacctg gtggagttca agaccatcta
     2341 catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat
     2401 cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca
     2461 ccacctgttc ctgtacggca tggacgagct gtacaagtaa gaattcgata tcaagcttat
     2521 cgataccgtc gacctcgagc ctctagaact atagtgagtc gtattacgta gatccagaca
     2581 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2641 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2701 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2761 ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct
     2821 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2881 ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg
     2941 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3001 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3061 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3121 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3181 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3241 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3301 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3361 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3421 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3481 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3541 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3601 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3661 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3721 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3781 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3841 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3901 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3961 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4021 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4081 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4141 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4201 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4261 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4321 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4381 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4441 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4501 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4561 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4621 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4681 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4741 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4801 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4861 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4921 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4981 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     5041 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     5101 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     5161 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     5221 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     5281 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5341 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5401 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5461 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     5521 tcgctattac gccagtcgac c
//
