LOCUS       Exported                6093 bp ds-DNA     circular SYN 18-APR-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    M3A_pTre2_2a_mCherry
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6093)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Jun 25, 2024 from SnapGene 2.8.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6093
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     protein_bind    15..285
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tetracycline response element"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        318..356
                     /note="minimal CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    507..2066
                     /note="Mex3a"
     misc_feature    930..1085
                     /note="KH domain"
     misc_feature    1179..1361
                     /note="KH domain"
     misc_feature    1911..2030
                     /note="RING Finger"
     CDS             2100..2153
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /note="T2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2160..2867
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /note="mCherry"
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     intron          2889..3461
                     /note="beta-globin intron"
                     /note="intron from rabbit beta-globin gene"
     polyA_site      3658..3713
                     /note="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal"
     rep_origin      complement(4275..4863)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5034..5894)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5895..5999)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 ctcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
       61 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
      121 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
      181 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
      241 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
      301 ctcggtaccc gggtcgaggt aggcgtgtac ggtgggaggc ctatataagc agagctcgtt
      361 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
      421 accgggaccg atccagcctc cgcggccccg aattcgagct cggtacccgg ggatcctcta
      481 gtcagctgac gcgtgctagc gcggccatgc ctagtctagt ggtatctgga ataatggaaa
      541 gaaatggggg ctttggagaa ctaggatgtt tcgggggaag cgctaaagac agggggcttc
      601 tggaagacga gcgcgccctc cagctggcac tggaccagct ctgccttctg gggctgggcg
      661 agccgcctgc acccaccgcc ggagaagatg gtgggggagg gggcggcgga gcaccagccc
      721 agcctgcagc accacctcag ccagctcccc ctcctcctcc agcagccgct cccgctgctc
      781 caagcgcagc ccctgccgct caaaccccac aacctcccac agcccctaag ggcgcaaacg
      841 atgccaagct gtgtgccctg tataaagagg cagaattgag gcttaaagga agctcaaaca
      901 ccacggagtg cgttcccgtg cccacctccg agcacgtggc cgagatcgtg ggcaggcaag
      961 gctgcaagat taaggctctg agggccaaaa ccaacaccta catcaagaca cctgtgcgag
     1021 gtgaggagcc agtgttcatg gtgactggga ggcgggaaga cgtggccaca gcccggcggg
     1081 agatcatctc tgcagctgag cacttctcca tgataagagc ctcacgcaac aagtctggcg
     1141 ccgcctttgg cgtggcccct gctctgccag gccaagtgac catccgtgta cgagtaccct
     1201 accgcgtggt gggactggtg gtgggcccca agggagcgac catcaaacgc atccagcagc
     1261 agaccaacac gtacatcatc actcccagcc gggaccgcga ccccgtgttt gagataaccg
     1321 gcgcaccggg caacgtggag cgtgcgcggg aggagatcga gacgcacatc gctgtgcgca
     1381 ccggcaagat tcttgaatac aacagcgacg gcgacttcct ggctggcagc cctgacgctg
     1441 ccttggacag ccgctactca gacgcctggc gggtgcatgc tccaggctgt aagccgctct
     1501 ccaccttcag gcagaacagc ctgggctgca taggcgaatg cggcgtggac tcggcctttg
     1561 aggccccgcg cctaagcgag cagggcgggg actttggcta cagcggatat ctgtttccgg
     1621 gctacggtgt gggcaagcag gacgtgtact atggcgtggc agagacaagc cccccgctgt
     1681 gggcgggtca ggagaatgcc acgcccacct cggtgctctt ctcctctgcc tcctcctcct
     1741 cctcttcagc caaggcacgc gcgggccccc caggcgcgca tcgctctcct gccacctctg
     1801 cggggccaga gcagctgacc gggctccctc ggaggccacc cggagagcca ctgcagggtt
     1861 tctctaaact ggggacaggc gggctgcgga gccccggcag cgggcgggac tgcatggtgt
     1921 gctttgagag tgaagtgaca gcagccctgg tgccctgtgg acacaacctt ttctgcatgg
     1981 agtgtgcagt acgcatctgc gagaggacgg acccagagtg tcctgtctgc cacatcaccg
     2041 ccacacaagc catccgaata ttttccggcc gcatcgataa gcttgtcgac ggcagtggag
     2101 agggcagagg aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccatctagag
     2161 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     2221 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2281 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     2341 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     2401 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     2461 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     2521 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     2581 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     2641 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     2701 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     2761 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     2821 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaatct agagctgaga
     2881 acttcagggt gagtttgggg acccttgatt gttctttctt tttcgctatt gtaaaattca
     2941 tgttatatgg agggggcaaa gttttcaggg tgttgtttag aatgggaaga tgtcccttgt
     3001 atcaccatgg accctcatga taattttgtt tctttcactt tctactctgt tgacaaccat
     3061 tgtctcctct tattttcttt tcattttctg taactttttt cgttaaactt tagcttgcat
     3121 ttgtaacgaa tttttaaatt cactttcgtt tatttgtcag attgtaagta ctttctctaa
     3181 tcactttttt ttcaaggcaa tcagggtaat tatattgtac ttcagcacag ttttagagaa
     3241 caattgttat aattaaatga taaggtagaa tatttctgca tataaattct ggctggcgtg
     3301 gaaatattct tattggtaga aacaactaca tcctggtaat catcctgcct ttctctttat
     3361 ggttacaatg atatacactg tttgagatga ggataaaata ctctgagtcc aaaccgggcc
     3421 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     3481 ttgttgtgct gtctcatcat tttggcaaag aattcactcc tcaggtgcag gctgcctatc
     3541 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     3601 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctgggtaat
     3661 aaaggaaatt tattttcatt gcaatagtgt gtgggaattt tttgtgtctc tcactcggaa
     3721 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     3781 caacatatgc catatgctgg ctgccatgaa caaaggtggc tataaagagg tcatcagtat
     3841 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     3901 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     3961 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     4021 cttctcttat gaactcgact gcattaatga atcggccaac gcgcggggag aggcggtttg
     4081 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4141 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4201 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4261 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4321 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4381 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4441 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     4501 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4561 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4621 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4681 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     4741 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4801 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4861 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4921 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4981 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5041 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5101 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5161 gcaatgatac cgcgagaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5221 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5281 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5341 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5401 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5461 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5521 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5581 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5641 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5701 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5761 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5821 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5881 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5941 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6001 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6061 tataaaaata ggcgtatcac gaggcccttt cgt
//
