LOCUS       62056_21625        6095 bp DNA     circular SYN 25-JAN-2021
DEFINITION  MCherry and eGFP expression vector pTTC-Dmel-WT, complete sequence.
ACCESSION   MN232102
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6095)
  AUTHORS   Boldyreva LV, Yarinich LA, Kozhevnikova EN, Ivankin AV, Lebedev MO, 
            Pindyurin AV.
  TITLE     Fine gene expression regulation by minor sequence variations 
            downstream of the polyadenylation signal
  JOURNAL   Mol Biol Rep (2021) In press
REFERENCE   2  (bases 1 to 6095)
  AUTHORS   Boldyreva LV, Ivankin AV, Kozhevnikova EN, Pindyurin AV.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUL-2019) Department of the Regulation of Genetic 
            Processes, Institute of Molecular and Cellular Biology, Lavrentyev 
            ave. 8/2, Novosibirsk, Novosibirsk Region 630090, Russia
REFERENCE   3  (bases 1 to 6095)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Biol 
            Rep (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-JUL-2019) Department of the Regulation of Genetic Processes, 
            Institute of Molecular and Cellular Biology, Lavrentyev ave. 8/2, 
            Novosibirsk, Novosibirsk Region 630090, Russia"
FEATURES             Location/Qualifiers
     source          1..6095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    838..859
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        874..904
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    912..928
                     /label=lac operator
                     /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)."
     primer_bind     936..952
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(989..1122)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1132..1836)
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     regulatory      complement(1898..2489)
                     /label=drosophila Hexokinase A (Hex-A) gene promoter
                     /note="drosophila Hexokinase A (Hex-A) gene promoter"
                     /regulatory_class="promoter"
     promoter        2496..2948
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     CDS             2959..3675
                     /label=EGFP
                     /note="enhanced GFP"
     regulatory      3771..3787
                     /label=human neuropilin 1 (NRP1) polyA signal
                     /note="human neuropilin 1 (NRP1) polyA signal"
                     /regulatory_class="polyA_signal_sequence"
     misc_feature    3794..3860
                     /note="piggyBac transposon 3' terminal repeat sequence
                     together with the adjacent TTAA target site"
     promoter        complement(3907..3925)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3946..3962)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3970..3986)
                     /label=lac operator
                     /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(3994..4024)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4039..4060)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4348..4936)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5110..5967)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5968..6072)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggggcgcgc
      661 cattctagat gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      721 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      781 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      841 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
      901 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
      961 cgccaagctc tagctagagg tcgacggtat acagacatga taagatacat tgatgagttt
     1021 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1081 attgctttat ttgtaaccat tataagctgc aataaacaag ttgatatctt acttgtacag
     1141 ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt
     1201 gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg cgccgggcag
     1261 ctgcacgggc ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc
     1321 cttcagcttc agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg
     1381 ctcggaggag gcctcccagc ccatggtctt cttctgcatt acggggccgt cggaggggaa
     1441 gttggtgccg cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg
     1501 ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc
     1561 ggggaaggac agcttcaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga
     1621 gccgtacatg aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt
     1681 caccttcagc ttggcggtct gggtgccctc gtaggggcgg ccctcgccct cgccctcgat
     1741 ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt
     1801 gatgatggcc atgttatcct cctcgccctt gctcacccca agctttcttc gtcgctgtct
     1861 ccgcttcttc ctgccataca tggatccgag ctcggtacca cacacacgat tctcgctctc
     1921 gccgtgtttt attttcgtcc gctgcacgcg tccaaacgct cggaacgttt tttacgaaat
     1981 taaattcact cgccgactgg tgctcacttt tcgctgtcgc tgctgctgcg ctgccggcgc
     2041 cgaccgcgac tgcgagccgc gctcacactc tcactctctc tctctctctc gctcgccgtg
     2101 aaggaacctt gaaggtcaaa aactgaaaat cggttttggg gcggcatggt tggagatatg
     2161 gcgtgtatat gtgcgacgct gaggggtcca caaatggcat attgataggg tgcgcgtgag
     2221 agagggggcg agagagtggc tatgaatgag aatgagtcaa acgcgtgttt atgcggaaac
     2281 ccttatgcgt cgcgtaagca atatcgtgat cacgcatttg ataaacattt taatttgtca
     2341 tctcaaatat agtcaatata tttcaggctt ttctcttcaa ggaaaaaaat tattttctta
     2401 agatgttaat ggtttaatta atgttaatgg ttaatggttt tgcatttctt tgacaaatta
     2461 aattggtcgc ctattatcac tgtgttaaag ctagcctaga atcccaaaac aaactggtta
     2521 ttgtggtagg tcatttgttt ggcagaaaga aaactcgaga aatttctctg gccgttattc
     2581 gttattctct cttttctttt tgggtctctc cctctctgca ctaatgctct ctcactctgt
     2641 cacacagtaa acggcatact gctctcgttg gttcgagaga gcgcgcctcg aatgttcgcg
     2701 aaaagagcgc cggagtataa atagaggcgc ttcgtctacg gagcgacaat tcaattcaaa
     2761 caagcaaagt gaacacgtcg ctaagcgaaa gctaagcaaa taaacaagcg cagctgaaca
     2821 agctaaacaa tctgcagtaa agtgcaagtt aaagtgaatc aattaaaagt aaccagcaac
     2881 caagtaaatc aactgcaact actgaaatct gccaagaagt aattattgaa tacaagaaga
     2941 gaactctgaa tagggaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     3001 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     3061 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     3121 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     3181 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3241 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3301 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3361 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     3421 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     3481 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3541 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     3601 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3661 gacgagctgt acaagtaaga attcgcggcc gcatacgatt taggtgacac tcgaggatcg
     3721 agatttaaat gagttgtggc cggcccttgt gactgggaaa accctggcgt aaataaaata
     3781 cgaaatgact agtcatgcgt caattttacg catgattatc tttaacgtac gtcacaatat
     3841 gattatcttt ctagggttaa tctagtatac gcgtatgcgg ccgcttaatt aatccagctt
     3901 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     3961 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4021 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4081 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4141 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4201 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4261 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4321 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4381 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4441 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4501 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4561 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4621 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4681 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4741 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4801 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4861 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4921 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4981 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5041 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5101 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5161 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5221 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5281 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5341 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5401 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5461 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5521 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5581 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5641 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5701 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5761 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5821 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5881 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5941 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6001 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6061 aggggttccg cgcacatttc cccgaaaagt gccac
//
