LOCUS       62056_21635        6199 bp DNA     circular SYN 25-JAN-2021
DEFINITION  MCherry and eGFP expression vector pTTC-Mmus-WT, complete sequence.
ACCESSION   MN232104
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6199)
  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 6199)
  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 6199)
  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..6199
                     /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)"
     promoter        complement(1913..1931)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1976..2179)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(2180..2483)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2554..3053
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3063..3779
                     /label=EGFP
                     /note="enhanced GFP"
     regulatory      3875..3891
                     /label=human neuropilin 1 (NRP1) polyA signal
                     /note="human neuropilin 1 (NRP1) polyA signal"
                     /regulatory_class="polyA_signal_sequence"
     misc_feature    3898..3964
                     /note="piggyBac transposon 3' terminal repeat sequence
                     together with the adjacent TTAA target site"
     promoter        complement(4011..4029)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4050..4066)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4074..4090)
                     /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(4098..4128)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4143..4164)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4452..5040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5214..6071)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6072..6176)
                     /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 agcgagggtc tccctatagt
     1921 gagtcgtatt aatttcgata agccagtaag cagtgggttc tctagttagc cagagagctc
     1981 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     2041 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     2101 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     2161 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     2221 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     2281 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     2341 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     2401 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     2461 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     2521 taattgatta ctattaatgc tagcaattct accgggtagg ggaggcgctt ttcccaaggc
     2581 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
     2641 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc
     2701 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg
     2761 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca
     2821 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
     2881 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg
     2941 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
     3001 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgcccggga
     3061 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     3121 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     3181 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     3241 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     3301 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     3361 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3421 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3481 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3541 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3601 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3661 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     3721 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     3781 aagaattcgc ggccgcatac gatttaggtg acactcgagg atcgagattt aaatgagttg
     3841 tggccggccc ttgtgactgg gaaaaccctg gcgtaaataa aatacgaaat gactagtcat
     3901 gcgtcaattt tacgcatgat tatctttaac gtacgtcaca atatgattat ctttctaggg
     3961 ttaatctagt atacgcgtat gcggccgctt aattaatcca gcttttgttc cctttagtga
     4021 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4081 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4141 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4201 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4261 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4321 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4381 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4441 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4501 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4561 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4621 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4681 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4741 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4801 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4861 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     4921 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4981 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5041 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5101 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5161 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5221 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5281 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5341 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5401 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5461 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5521 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5581 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5641 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5701 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5761 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5821 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5881 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5941 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6001 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6061 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6121 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6181 tttccccgaa aagtgccac
//
